Disulfide derivatives useful for treating allergic diseases

ABSTRACT

Novel disulfide derivatives useful for preventing or treating allergic diseases of the eye, nose, skin, ear, gastrointestinal tract, airways or lung and preventing or treating manifestations of systemic mastocytosis are disclosed. The disulfide derivatives act as mast cell stabilizers.

[0001] This application claims priority from co-pending U.S. Provisional Application, U.S. Serial No. 60/205,827, filed May 19, 2000.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention relates to novel disulfide derivatives useful for treating allergic diseases.

[0004] 2. Description of the Related Art

[0005] Antihistamines and mast cell stabilizers are two types of drugs currently used topically to treat allergic diseases. Antihistamine drugs are used to interrupt the allergic effects that histamine causes after it has been released from a mast cell. Many topical antihistamine drugs are marketed. For example, emedastine difumarate and levocabastine hydrochloride are available for ocular allergies (see Ophthalmic Drug Facts 1999, Facts and Comparisons, St. Louis, Mo., pp. 59-80).

[0006] Mast cell stabilizers prevent mast cells from “degranulating” or releasing histamine and other components or “mediators” during an allergic reaction. Examples of ophthalmic drugs marketed as mast cell stabilizers include olopatadine (see U.S. Pat. No. 5,641,805) and cromolyn sodium.

[0007] U.S. Pat. No. 4,705,805 discloses certain disulfide derivatives that are useful as anti-thrombotic agents. The disulfide derivatives suppress blood platelet aggregation. The '705 patent does not disclose the use of disulfide derivatives in the topical treatment of allergic diseases of the eye or nose.

SUMMARY OF THE INVENTION

[0008] The present invention provides methods for preventing or treating allergic diseases of the eye, nose, skin, ear, gastrointestinal tract, airways or lung. The methods may also be used to treat manifestations of systemic mastocytosis. The methods of the present invention comprise topically or systemically administering to a patient a novel mast cell stabilizing disulfide derivative of the formula

[0009] wherein

[0010] X=—NHC(═O)NH—R;

[0011] R=H; (un)substituted phenyl; (un)substituted benzyl; or C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R²;

[0012] R²=C₁-C₃ alkyl; and

[0013] R³ and R⁴ are independently H; benzyl; C₁-C₈ alkyl or alkenyl; C₄-C₇ cycloalkyl; (un)substituted aryl; or (un)substituted 5-7 membered heterocyclic ring; wherein optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².

[0014] The present invention is also directed toward topically or systemically administrable compositions for treating or preventing allergic diseases of the eye, nose, skin, ear, gastrointestinal tract, airways or lung and treating or preventing manifestations of systemic mastocytosis, wherein the compositions comprise a disulfide derivative of formula (I).

DETAILED DESCRIPTION OF THE INVENTION

[0015] The disulfide derivatives of formula (I) can be made as described in Scheme 1.

[0016] The appropriate isocyanate is added to a stirring solution of the bis amino-disulfide in a solvent such as tetrahydrofuran or methylene chloride at a temperature between −20° C. and 30° C. An organic base such as triethylamine, or pyridine is added after the reaction mixture has stirred for 5 to 30 minutes and the reaction is stirred for 6 to 48 hr. The disulfides can then be isolated using standard, known procedures.

[0017] Preferred compounds of formula (I) are those having the X substituents in the ortho position and wherein R=C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R². Most preferred are compounds wherein R=C₁-C₅ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².

[0018] Compounds of formula (I) may be administered topically (i.e., local, organ-specific delivery) or systemically by means of conventional topical or systemic formulations, such as solutions, suspensions or gels for the eye and ear; nasal sprays or mists for the nose; metered dose inhalers for the lung; solutions, gels, creams or lotions for the skin; oral dosage forms including tablets or syrups for the gastrointestinal tract; and parenteral dosage forms including injectable formulations. The concentration of the compound of formula (I) in the formulations of the present invention will depend on the selected route of administration and dosage form. The concentration of the compound of formula (I) in topically administrable formulations will generally be about 0.00001 to 5 wt. %. For systemically administrable dosage forms, the concentration of the compound of formula (I) will generally range from about 10 mg to 1000 mg.

[0019] The preferred formulation for topical ophthalmic administration is a solution intended to be administered as eye drops. For solutions intended for topical administration to the eye, the concentration of the compound of formula (I) is preferably 0.0001 to 0.2 wt. %, and most preferably from about 0.0001 to 0.01 wt. %. The topical compositions of the present invention are prepared according to conventional techniques and contain conventional excipients in addition to one or more compounds of formula (I). A general method of preparing eye drop compositions is described below:

[0020] One or more compounds of formula (I) and a tonicity-adjusting agent are added to sterilized purified water and if desired or required, one or more excipients. The tonicity-adjusting agent is present in an amount sufficient to cause the final composition to have an ophthalmically acceptable osmolality (generally about 150-450 mOsm, preferably 250-350 mOsm). Conventional excipients include preservatives, buffering agents, chelating agents or stabilizers, viscosity-enhancing agents and others. The chosen ingredients are mixed until homogeneous. After the solution is mixed, pH is adjusted (typically with NaOH or HCl) to be within a range suitable for topical ophthalmic use, preferably within the range of 4.5 to 8.

[0021] Many ophthalmically acceptable excipients are known, including, for example, sodium chloride, mannitol, glycerin or the like as a tonicity-adjusting agent; benzalkonium chloride, polyquaternium-1 or the like as a preservative; sodium hydrogenphosphate, sodium dihydrogenphosphate, boric acid or the like as a buffering agent; edetate disodium or the like as a chelating agent or stabilizer; polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid, polysaccharide or the like as a viscosity-enhancing agent; and sodium hydroxide, hydrochloric acid or the like as a pH controller.

[0022] If required or desired, other drugs can be combined with the disulfide derivatives of formula (I), including, but not limited to, antihistaminic agents, anti-inflammatory agents (steroidal and non-steroidal), and decongestants. Suitable antihistaminic agents include emedastine, mapinastine, epinastine, levocabastine, loratadine, desloratadine, ketotifen, azelastine, cetirazine, and fexofenadine. The preferred antihistaminic agent for ophthalmic use is emedastine, which is generally included in topically administrable compositions at a concentration of 0.001-0.1 wt. %, preferably 0.05 wt. %. Suitable anti-inflammatory agents includemometasone, fluticasone, dexamethasone, prednisolone, hydrocortisone, rimexolone and loteprednol. Suitable decongestants include oxymetazoline, naphazoline, tetrahydrozoline, xylometazoline, propylhexedrine, ethyinorepinephrine, pseudoephedrine, and phenylpropanolamine.

[0023] According to the present invention, the disulfide derivatives of formula (I) are useful for preventing and treating ophthalmic allergic disorders, including allergic conjunctivitis, vernal conjunctivitis, vernal keratoconjunctivitis, and giant papillary conjunctivitis; nasal allergic disorders, including allergic rhinitis and sinusitis; otic allergic disorders, including eustachian tube itching; allergic disorders of the upper and lower airways, including intrinsic and extrinsic asthma; allergic disorders of the skin, including dermatitis, eczema and urticaria; allergic disorders of the gastrointestinal tract, including systemic anaphylaxis resulting from ingestion of allergen and iatrogenic anaphylaxis caused by contrast agents used during diagnostic imaging procedures; and manifestations of systemic mastocytosis including hypotension.

[0024] The following examples are intended to be illustrative but not limiting.

[0025] EXAMPLE 1 Topical Ophthalmic Solution Formulation Ingredient Concentration (wt. %) Compound of formula (I) 0.0001 to 0.2 Dibasic Sodium Phosphate (Anhydrous) 0.5 Sodium Chloride 0.65 Benzalkonium Chloride 0.01 NaOH/HCl q.s. pH 6-8 Purified Water q.s. 100

[0026] EXAMPLE 2 Topical Ophthalmic Gel Formulation Ingredient Concentration (wt. %) Compound of formula (I) 0.0001 to 0.2 Carbopol 974 P 0.8 Edetate Disodium 0.01 Polysorbate 80 0.05 Benzalkonium Chloride 0.01 NaOH/HCl q.s. pH 6-8 Water for Injection q.s. 100

EXAMPLE 3 Synthesis of bis-[2-(3-allylurea)-phenyl)]-disulfide (II)

[0027] To a stirred solution of 2-aminophenyl disulfide (1.5 g, 6 mmol) in 10 ml of THF, was added allyl isocyanate (1.1 ml, 12 mmol). After stirring at room temperature for 5 min, 1 ml of triethylamine was added. The resulting mixture was stirred and refluxed for 18 hr. After cooling, the solvent was evaporated and the solids were filtered off. The filtrate was washed with 5% of HCl, saturated NaHCO₃ and saturated NaCl and then dried over MgSO₄. Concentration under reduced pressure and chromatography of the residue on silica gel, eluting with 30% of ethyl acetate in hexane to 60% of ethyl acetate in hexane gave 0.31 g of 11 as a white solid.

[0028]¹H NMR (CDCl₃) δ 8.14-7.90 (m, 4H), 7.33-6.86 (m, 8H), 5.95-5.79 (m, 2H), 5.25-5.07 (m, 4H), 3.33-3.31 (m, 4H). ¹³C NMR (CDCl₃) δ 154.94 (C═O), 140.28 (C), 135.95 (CH), 133.51 (CH), 130.04 (CH), 124.91 (C), 122.45 (CH), 121.46 (CH), 114.91 (CH₂), 41.93 (CH₂). Analysis calculated for C₂₀H₂₂O₂N₄S₂ requires: C, 57.95; H, 5.35; N, 13.52%. Found: C, 57.91; H, 5.39; N, 13.46%.

EXAMPLE 4 Mast Cell Activity

[0029] Preparation of Cell Suspension

[0030] Methods detailing preparation of monodispersed HCTMC and mediator release studies with these cells have been described (U.S. Pat. No. 5,360,720 and Miller et al, Ocular Immunology and Inflammation, 4(1):39-49 (1996)). Briefly, human conjunctival tissue mast cells were isolated from post-mortem tissue donors obtained within 8 hours of death by various eye banks and transported in Dexsol® corneal preservation medium, or equivalent. Tissues were enzymatically digested by repeated exposure (30 min. at 37° C.) to collagenase and hyaluronidase (2× with 200 U each/gram tissue, then 24× with 2000 U each/gram tissue) in Tyrode's buffer containing 0.1% gelatin. (Tyrode's buffer (in mM): 137 NaCl, 2.7 KCl, 0.35 NaH₂PO4, 1.8 CaCl₂, 0.98 MgCl₂, 11.9 NaHCO₃, and 5.5 glucose). Each digestion mixture was filtered over Nitex® cloth (100 μm mesh, Tetko, Briarcliff Manor, N.Y.) and washed with an equal volume of buffer. Filtrates were centrifuged at 825× g (7 min). Pellets were resuspended in buffer then combined for enrichment over a 1.058 g/L Percoll® cushion. The enriched pellet was washed, resuspended in supplemented RPMI 1640 medium and incubated at 37° C. to equilibrate.

[0031] Histamine Release Studies

[0032] Cells were harvested from the culture plate and counted for viability (trypan blue exclusion) and mast cell number (toluidine blue O). Mast cells (5000/tube; 1 mL final volume) were challenged (37° C.) for 15 min with goat-anti-human IgE (10 μg/mL) following treatment (15 minutes; 37° C.) with test drug or Tyrode's buffer. Total and non-specific release controls were exposed to 0.1% Triton X-100 and goat IgG (10 μg/mL), respectively. The reaction was terminated by centrifugation (500× g, 4° C., 10 min). Supernatants were stored at −20° C. until analyzed for histamine content by RIA (Beckman Coulter, Chicago, Ill.).

[0033] Preparation of Test Drug Solutions

[0034] All test drugs were made to solution immediately prior to use. Each was dissolved in DMSO at 10 mM or greater concentration and then diluted in Tyrode's buffer containing 0.1% gelatin over the concentration for evaluation.

[0035] Data Analysis

[0036] Inhibition of histamine release was determined by direct comparison of with anti-IgE challenged mast cells using Dunnett's t-test (Dunnett, “A multiple comparison procedure for comparing treatments with a control”, J. Amer. Stat. Assoc. (1955), 50:1096-1121). An IC50 value (the concentration at which the test compound inhibits histamine release at a level of 50% compared to the positive control) was determined by 4-parameter logistic fitting using the Levenburg-Marquardt algorithm or by linear regression. The results are reported in Table 1. TABLE 1

COMPOUND NO. T X MOLSTRUCTURE IC50(nM) 1 S—S

314  10 175 278 [194]

[0037] The data shown in Table 1 indicate that the compounds of formula (I) potently inhibit histamine release from human conjunctival mast cells in an in vitro model of allergic conjunctivitis. EXAMPLE 5 Topical Ophthalmic Solution Formulation Ingredient Concentration (wt. %) Compound of formula (I) 0.0001 to 0.2 Emedastine 0.001 to 0.1 Dibasic Sodium Phosphate (Anhydrous) 0.5 Sodium Chloride 0.65 Benzalkonium Chloride 0.01 NaOH/HCl q.s. pH 6-8 Purified Water q.s. 100

[0038] The invention has been described by reference to certain preferred embodiments; however, it should be understood that it may be embodied in other specific forms or variations thereof without departing from its spirit or essential characteristics. The embodiments described above are therefore considered to be illustrative in all respects and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description. 

What is claimed is:
 1. A disulfide derivative of the formula

wherein X=—NHC(═O)NH—R; R=H; (un)substituted phenyl; (un)substituted benzyl; or C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R²; R²=C₁-C₃ alkyl; and R³ and R⁴ are independently H; benzyl; C₁-C₈ alkyl or alkenyl; C₄-C₇ cycloalkyl; (un)substituted aryl; or (un)substituted 5-7 membered heterocyclic ring; wherein optional substituents are selected from the group 2 consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 2. The disulfide derivative of claim 1 wherein the X substituents are in the ortho position and wherein R=C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 3. The disulfide derivative of claim 2 wherein R=C₁-C₅ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 4. The disulfide derivative of claim 3 wherein X is —NHC(═O)NH—CH₂—CH═CH₂.
 5. A topically or locally administrable pharmaceutical composition for treating allergic diseases of the eye, nose, skin, ear or lung comprising a tonicity-adjusting agent in an amount sufficient to cause the composition to have an osmolality of 150-450 mOsm, a pharmaceutically acceptable preservative and a disulfide derivative of the formula

wherein X=—NHC(═O)NH—R; R=H; (un)substituted phenyl; (un)substituted benzyl; or C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R²; R²=C₁-C₃alkyl; and R³and R⁴ are independently H; benzyl; C₁-C₈ alkyl or alkenyl; C₄-C₇ cycloalkyl; (un)substituted aryl; or (un)substituted 5-7 membered heterocyclic ring; wherein optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 6. The pharmaceutical composition of claim 5 wherein the X substituents are in the ortho position and wherein R=C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 7. The pharmaceutical composition of claim 6 wherein R=C₁-C₅ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 8. The pharmaceutical composition of claim 7 wherein X is —NHC(═O)NH—CH₂—CH═CH₂.
 9. The pharmaceutical composition of claim 5 wherein the disulfide derivative is present in an amount from about 0.00001 to 5 wt. %.
 10. The pharmaceutical composition of claim 9 wherein the disulfide derivative is present in an amount from about 0.0001 to 0.2 wt. %.
 11. The pharmaceutical composition of claim 10 wherein the disulfide derivative is present in an amount from about 0.0001 to 0.01 wt. %.
 12. A method for preventing or treating an allergic disease of the eye, nose, skin, ear, gastrointestinal tract or lung and preventing or treating manifestations of systemic mastocytosis in a patient comprising administering to the patient a composition comprising a disulfide derivative of the formula

wherein X=—NHC(═O)NH—R; R=H; (un)substituted phenyl; (un)substituted benzyl; or C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R²; R²=C₁-C₃ alkyl; and R³ and R⁴ are independently H; benzyl; C₁-C₈ alkyl or alkenyl; C₄-C₇ cycloalkyl; (un)substituted aryl; or (un)substituted 5-7 membered heterocyclic ring; wherein optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 13. The method of claim 12 wherein the X substituents are in the ortho position and wherein R=C₁-C₈ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 14. The method of claim 13 wherein R=C₁-C₅ alkyl or alkenyl, optionally substituted with or terminated by OH, OR², NR³R⁴; C₄-C₇ cycloalkyl, (un)substituted aryl, or (un)substituted 5-7 membered heterocyclic ring; where optional substituents are selected from the group consisting of C₁-C₆ alkyl or alkoxy; halogen; OH; CN; CF₃; NO₂; and CO₂R².
 15. The method of claim 14 wherein X is —NHC(═O)NH—CH₂—CH═CH₂.
 16. The method of claim 12 wherein the disulfide derivative is administered to the patient in a topically administrable composition containing the disulfide derivative in an amount from about 0.00001 to 5 wt. %.
 17. The method of claim 16 wherein the disulfide derivative is present in an amount from about 0.0001 to 0.2 wt. %.
 18. The method of claim 17 wherein the disulfide derivative is present in an amount from about 0.0001 to 0.01 wt. %.
 19. The method of claim 12 wherein the disulfide derivative is administered to the patient in a systemically administrable composition containing the disulfide derivative in an amount from about 10 to 1000 mg. 